ATCC has successfully irradiated (SCRC-1040.1) and treated the cells with Mitomycin C (SCRC-1040.2a) for use as a feeder layer.

The cells can be used as a feeder layer to support the growth of embryonic stem (ES) cells and for the maintenance of ES cells in the undifferentiated state.

Storage Conditions

liquid nitrogen vapor phase

Derivation

The cell line was established by ATCC in 2003 from embryonic day 14 (E14) CF-1 mouse embryos.

Clinical Data

male and female mixed

Comments

The growth of these cells should be arrested before being used as a feeder layer. ATCC has successfully irradiated (SCRC-1040.1™) and treated the cells with Mitomycin C (SCRC-1040.2a™) for use as a feeder layer. If the MEFs are being used as a feeder layer for ES cells, it is not recommended to use them past passage no. 6 (P6).

Complete Growth Medium

The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:

fetal bovine serum to a final concentration of 15%

This medium is formulated for use with a 5% CO2 in air atmosphere. (Standard DMEM formulations contain 3.7 g/L sodium bicarbonate and a 10% CO2 in air atmosphere is then recommended).

Subculturing

To insure the highest level of viability, be sure to warm media and Trypsin / EDTA to 37ºC before using it on the cells. Cells should be split when they reach confluency. Split cells at approximately 0.4 X 104 cells/cm2

Remove and discard culture medium.

Briefly rinse the cell layer with 1XPBS (SCRR-2201) solution to remove all traces of serum, which contain trypsin inhibitor.

Freshney RI. Culture of Animal Cells: A Manual of Basic Technique, 5th edition. New York: Wiley Liss; 2005. For more information on enzymatic dissociation and subculturing of cell lines see Chapter 13.

Permits

These permits may be required for shipping this product:

Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.